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Team:Paris Saclay/Notebook/August/4
From 2014.igem.org
Contents |
Monday 4th August
Lab Work
A - The chassis coli Odor free
Striate on Dishes
by Juliette & Romain
We striated MG1655 and MG1655Z1 on LB and Kan dishes, from the striates prepared on LB dishes 1st August
C - Salicylate Inducible Suppressing System
Plasmid DNA Purification
by Eugene and Fabio
- Ligation of BBa_J61051 and BBa_K228001 made the 30th July
From Liquid Culture made the 1st August
Digestion to check
In progress
Electrophoresis
In progress
D - Lemon scent
Gel electrophoresis of BBa_K762100
by Sean
Samples used were prepared on the 30th July.
Results
From left to right: 5 µl of BBa_K762100 + "old" Phusion enzyme, 5 µl of BBa_K762100 + "new" Phusion enzyme
Electroporation
by Terry
Strain used: DY330 containing plasmid pJBEI6409.
We had to switch the Limonen Synthase cassette with the Apramycin resistance.
The culture has been placed at 42°C for 17 min then cooled 10 min before centrifugation.
Polymerase chain reaction
by Sean & Pierre
BBa_K517003
| component | volume |
|---|---|
| H2O | 35.5μl |
| Phusion buffer 5X | 10μl |
| dNTPs | 2μl |
| iPS68bis | 1μl |
| iPS69 | 1μl |
| DNA | 1μl |
| Phusion enzyme | 0.5μl |
BBa_K762100
For this BioBrick two tubes were prepared: one with undiluted DNA and one with DNA diluted 10-1.
| component | volume |
|---|---|
| H2O | 35.5μl |
| Phusion buffer 5X | 10μl |
| dNTPs | 2μl |
| iPS66 | 1μl |
| iPS67 | 1μl |
| DNA | 1μl |
| Phusion enzyme | 0.5μl |
Tubes were placed in PCR machine with the following parameters.
| Cycle step | Temperature | Time | Cycle |
|---|---|---|---|
|
Initial denaturation |
98°C |
1 min |
1 |
| Denaturation | 98°C | 15 s | 25 - 30 |
| Annealing | 52°C | 25 s | 25 - 30 |
| Extension | 72°C | 45 s | 25-30 |
| Final extension | 72°C | 10 min | 1 |
| Final extension | 8°C | hold | 1 |
Miscellaneous
By Hoang Vu
Ethics
Well, I just got back from one month of vacation and when I came, everybody was already on lab works. So, I decided to work on ethics first. Synthetic Biology and BioArt are controversial because one is about engineering living organisms, and therefore raises economic, religious, political, moral problems; and the other is about engineering the living organisms too but it's mostly USING the living organisms to do art, to carry a message. This is how I see the essential points of our project ethical problems. But my point of view is not fixed. Maybe I'm wrong, maybe there is something that I have missed. That's why I started with a research on GMOs in general to better understand those issues. - The first trangenic pet were GloFish, "TK-1" 2003 in Taiwan - California is completely against such animals but the funny thing is that California is one of the best supporters of agricultural GMOs. So, there is the question: Why the plants and not the fish? Probably economic benefits and ethical positions are the reasons. Then, is there a hierachy of living organisms? Why? - GMOs under commercial cover raise new problems on environment and commercial rights. - Take a look at the arguments between Anti-GMO and Pro-GMO movements. The 90's context for example in Europe after food crises like the mad cow disease, contribute to the incomplete acceptance by the Europeans.
Bibliography
I worked on ethics till one of our advisors ask me: "Can you do a research on the chemical components of the lemon odor please? I need more publications on chemical characterization of limonene, citral A and B, and beta-Pinene"
Members there:
- Instructors and advisors: Alice, Solenne and Sylvie.
- Students: Eugene, Fabio, Hoang Vu, Juliette, Pierre, Romain, Sean and Terry.
